RESEARCH ARTICLE
Multiplexing detection of IgG against Plasmodium falciparum pregnancy-specific antigens Ana Maria Fonseca1,2, Llorenc¸ Quinto1, Alfons Jime´nez1,3, Raquel Gonza´lez1,4, Azucena Bardajı´1,4, Sonia Maculuve4, Carlota Dobaño1,4, Maria Rupe´rez1,4, Anifa Vala4, John J. Aponte1,4, Esperanza Sevene4,5, Eusebio Macete4, Clara Mene´ndez1,4, Alfredo Mayor1,4*
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1 ISGlobal, Barcelona Ctr. Int. Health Res. (CRESIB), Hospital Clı´nic - Universitat de Barcelona, Barcelona, Spain, 2 Graduate Program in Areas of Basic and Applied Biology (GABBA), Universidade do Porto, Porto, Portugal, 3 Spanish Consortium for Research in Epidemiology and Public Health (CIBERESP), Madrid, Spain, 4 Centro de Investigac¸ão em Sau´de da Manhic¸a (CISM), Maputo, Mozambique, 5 Eduardo Mondlane University, Maputo, Mozambique *
[email protected]
Abstract OPEN ACCESS Citation: Fonseca AM, Quinto L, Jime´nez A, Gonza´lez R, Bardajı´ A, Maculuve S, et al. (2017) Multiplexing detection of IgG against Plasmodium falciparum pregnancy-specific antigens. PLoS ONE 12(7): e0181150. https://doi.org/10.1371/journal. pone.0181150 Editor: Takafumi Tsuboi, Ehime Daigaku, JAPAN Received: February 28, 2017
Background Pregnant women exposed to Plasmodium falciparum generate antibodies against VAR2CSA, the parasite protein that mediates adhesion of infected erythrocytes to the placenta. There is a need of high-throughput tools to determine the fine specificity of these antibodies that can be used to identify immune correlates of protection and exposure. Here we aimed at developing a multiplex-immunoassay to detect antibodies against VAR2CSA antigens.
Accepted: June 27, 2017 Published: July 17, 2017
Methods and findings
Copyright: © 2017 Fonseca et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
We constructed two multiplex-bead arrays, one composed of 3 VAR2CSA recombinantdomains (DBL3X, DBL5Ɛ and DBL6Ɛ) and another composed of 46 new peptides covering VAR2CSA conserved and semi-conserved regions. IgG reactivity was similar in multiplexed and singleplexed determinations (Pearson correlation, protein array: R2 = 0.99 and peptide array: R2 = 0.87). IgG recognition of 25 out of 46 peptides and all recombinant-domains was higher in pregnant Mozambican women (n = 106) than in Mozambican men (n = 102) and Spanish individuals (n = 101; p
